Rapid detection of porcine reproductive and respiratory syndrome virus by reverse transcription loop-mediated isothermal amplification assay.
A rapid detection assay based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) has been developed for detecting porcine reproductive and respiratory syndrome virus (PRRSV). The RT-LAMP assay utilized a set of six primers to amplify the open reading frame 6 (ORF6) of the PRRSV. The amplified products were analyzed by agarose gel electrophoresis or visualized by colorimetric method. The results demonstrated that the RT-LAMP assay detected all 22 different PRRSV isolates, had no cross-reaction with four other swine viruses (i.e., PCV2, SIV, CSFV, and PEDV), and obtained a 91.3% sensitivity in 23 positive clinical samples in reference to the permissive cells-based virus isolation procedure. Therefore, the RT-LAMP assay provides a specific and sensitive means for detecting PRRSV in a simple, fast, and cost-effective manner. Furthermore, the RT-LAMP assay can be performed in less well-equipped laboratories as well as fields.
Li
Qiang
Q
State Key Laboratory of Biocontrol, College of Life Sciences, Sun Yat-Sen University, Guangzhou 510006, China.
Zhou
Qing-Feng
QF
Xue
Chun-Yi
CY
Ma
Jing-Yun
JY
Zhu
Dao-Zhong
DZ
Cao
Yong-Chang
YC
D000818:Animals; D002460:Cell Line; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis / Q000821:virology*; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013552:Swine; D013997:Time Factors